In vivo cloning of defined chromosomal segments into YAC vectors

该项目寻求开发一种技术，在要克隆的基因已经被映射到生物体的RFLP图谱中的两个限制性片段标记之间的情况下，将绕过基因组长距离图谱行走的需要。可克隆区域的大小不会由体外克隆的通常限制决定，因为这一过程将涉及到拟南芥染色体和专门的酵母人工染色体(YAC)载体之间的体内定向基因重组。其核心思想是通过细胞融合和体内同源重组，在一步内将确定的大物理尺寸的遗传区间克隆到YAC载体中。这项技术的发展将提高以拟南芥为模型的科学家克隆整个基因组的能力，所需时间比现在预测的要短得多。

This project seeks to develop technology that will bypass the need for genomic walking over long map distances in those cases where the gene to be cloned has been mapped between two restriction fragment markers in an RFLP map of the organism. The size limit of the clonable region will not be determined by the usual limitations of in vitro cloning because the procedure will involove targeted in vivo genetic recombination between Arabidopsis chromosomes and specialized Yeast Artifical Chromosome (YAC) vectors. The central idea is to clone a defined genetic interval of large physical size in a single step into a YAC vector by cell fusion and in vivo homologous recombination. %%% The development of this technology will enhance the ability of scientists using Arabidopsis as a model to clone the entire genome in much less time than is now predicted.